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Briefly, HaCaT keratinocytes were seeded at a density of 10,000 cells per well in 200 l fresh culture medium (DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin solution) into 96-well flat-bottomed microtitre plates (Costar, Corning, USA) and incubated at 37 C in humidified 5% CO 2 using AutoFlow NU-5510 Direct Heat CO 2 Incubator (NuAire, USA) for 24 hours before treatment
Fresh combinations prevent unexpected interactions between compounds
The dose lands in muscle tissue and is taken up into the bloodstream over the following hours
1 mL syringe required BAC water added 2.5 mL Final concentration 200 mg/mL 25 mg draw 0.125 mL / 12.5 units
